purmorphamine促进人牙周膜干细胞应力成骨

Purmorphamine promotes osteogenic differentiation in human periodontal ligament stem cells under dynamic tensile

目的研究purmorphamine在动态张应力促人牙周膜干细胞(human periodontal ligament stem cells,PDLSCs)向成骨细胞分化过程中的作用。方法分离培养鉴定PDLSCs,在矿化诱导环境中加入purmorphamine,采用Flexcell FX-4000T应力加载系统对细胞加力24 h,以Real-time PCR检测成骨相关指标Runx2、alkaline phosphatase(ALP)以及Hedgehog(Hh)通路的标志物GLI1、Pathed1(PTCH1)、Smoothend(SMO)。结果动态张应力作用24 h后,成骨相关指标Runx2、ALP,Hh通路的标志物GLI1、PTCH1、SMO的表达水平明显升高(P<0.05);加入purmorphamine后,成骨相关指标Runx2、ALP,Hh通路的标志物GLI1、PTCH1、SMO的表达水平较加力组均有增强,差异有统计学意义(P<0.05)。结论 purmor-phamine可能通过激活Hh通路,进而增强人PDLSCs应力条件下向成骨细胞分化。

Objective To determine the effect of purmorphamine on the osteogenic differentiation in human periodontal ligament stem cells（PDLSCs） under dynamic strain.Methods PDLSCs were isolated from freshly extracted teeth and identified with flow cytometry for expression of STRO-1 and CD146,and then subcultured into six-well flexible-bottomed Uniflex culture plates.When 80% confluence was achieved,purmorphamine,an activator of the hedgehog（Hh） signaling pathway,was added into the wells in mineralization induction medium at the concentration of 1 μmol/L,and dynamic strains were applied to PDLSCs with the Flexcell FX-4000T Tension Plus System for 24 h.Then the mRNA expression of osteoblastic related indexes of Runx2,ALP and Hh pathway markers GLI1,PTCH1,SMO was detected by real-time PCR.Results After 24 h loading,the gene expression of osteogenic markers Runx2 and ALP and Hh pathway markers（GLI1,PTCH1 and SMO） was all increased（P0.05）.And the group with purmorphamine treatment showed much higher expression of these genes than the control group and the only mechanical loading group（P0.05）.Conclusion Purmorphamine promotes osteogenic differentiation in PDLSCs under dynamic tensile,probably through activation of Hh signal pathway.